Limbacher Melissa R, Puglia Megan K, Riccardi Caterina M, Kumar Challa V
Department of Chemistry, University of Connecticut, Storrs, CT, United States.
Department of Chemistry, University of Connecticut, Storrs, CT, United States; Department of Molecular and Cell Biology, University of Connecticut, Storrs, CT, United States.
Methods Enzymol. 2018;609:1-22. doi: 10.1016/bs.mie.2018.06.012. Epub 2018 Aug 28.
A simple method for interlocking glucose oxidase and horseradish peroxidase in a network of cellulose fibers coated with bovine serum albumin (BSA)-exfoliated graphene (biographene) is reported here. The resulting paper reactor is inexpensive and stable. Biographene is expected to function as an electron shuttle, making the reaction between the enzyme and the substrate more efficient, and this hypothesis is examined here. The BSA used to separate the sheets of graphene provides extra carboxylic acid groups and primary amines to help interlock the enzymes and the graphene in between the fibers. The decrease in entropy associated with interlocking the enzymes on a solid support is likely responsible for the increase in enzymatic stability/activity observed. Each cellulose disk contained 5.2mg of enzyme per gram of paper and 93% of the enzyme is retained after washing for 0.5-2h. This simple methodology provides a low cost, effective approach for achieving high enzymatic activity and good loadings on a benign, versatile support.
本文报道了一种在涂有牛血清白蛋白（BSA）-剥离石墨烯（生物石墨烯）的纤维素纤维网络中使葡萄糖氧化酶和辣根过氧化物酶相互锁定的简单方法。所得的纸质反应器价格低廉且稳定。生物石墨烯有望作为电子穿梭体，使酶与底物之间的反应更高效，本文对这一假设进行了检验。用于分离石墨烯片层的BSA提供了额外的羧酸基团和伯胺，有助于使酶与纤维间的石墨烯相互锁定。与在固体载体上锁定酶相关的熵的降低可能是观察到的酶稳定性/活性增加的原因。每个纤维素圆盘每克纸含有(5.2)毫克酶，洗涤(0.5 - 2)小时后(93%)的酶得以保留。这种简单的方法为在良性、通用的载体上实现高酶活性和良好负载量提供了一种低成本、有效的途径。
